Makino S, Harada M, Akashi K, Taniguchi S, Shibuya T, Inaba S, Niho Y
Blood Transfusion Service, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Bone Marrow Transplant. 1991 Oct;8(4):239-44.
A simplified method was established for cryopreservation of peripheral blood stem cells (PBSCs) using hydroxyethyl starch (HES) and dimethylsulfoxide (DMSO) as a cryoprotective agent at -80 degrees C without rate-controlled freezing. The data indicate that a cryoprotective solution consisting of 6% HES and 5% DMSO produced the highest recovery rates for nucleated cells (92.0 +/- 3.5%), CFU-GM (73.8 +/- 4.1%) and BFU-E (82.2 +/- 6.9%), and the highest trypan blue viability (88.4 +/- 3.6%). For long-term cryopreservation of PBSCs, CFU-GM recovery rates remained almost unchanged during 5-18 months; the mean CFU-GM recovery rate after 18 months of cryopreservation was 70 +/- 11%. When large-scale samples of PBSCs in 100-ml freezing bags were cryopreserved for clinical use, cell and CFU-GM recoveries were similar. Using this method, 10 patients with hematological malignancy received PBSC transplants after marrow-ablative chemotherapy. All demonstrated early engraftment; seven are now alive in unmaintained complete remission for 3.5-15 months after PBSC transplant. This simple and inexpensive method will be useful for the wider application of PBSC transplant as a therapeutic alternative in the treatment of malignant diseases.
建立了一种简化方法，使用羟乙基淀粉（HES）和二甲基亚砜（DMSO）作为冷冻保护剂，在-80℃下对外周血干细胞（PBSCs）进行无速率控制冷冻保存。数据表明，由6% HES和5% DMSO组成的冷冻保护溶液对有核细胞（92.0±3.5%）、CFU-GM（73.8±4.1%）和BFU-E（82.2±6.9%）产生了最高的回收率，以及最高的台盼蓝活力（88.4±3.6%）。对于PBSCs的长期冷冻保存，CFU-GM回收率在5至18个月期间几乎保持不变；冷冻保存18个月后的平均CFU-GM回收率为70±11%。当在100毫升冷冻袋中对大规模PBSCs样本进行临床冷冻保存时，细胞和CFU-GM回收率相似。使用这种方法，10例血液系统恶性肿瘤患者在骨髓清除化疗后接受了PBSC移植。所有患者均表现出早期植入；7例患者在PBSC移植后3.5至15个月处于未维持的完全缓解状态且存活。这种简单且成本低廉的方法将有助于PBSC移植作为恶性疾病治疗的一种治疗选择更广泛地应用。
